LMS cells were grown as previously described [28 (link)]. Primary antibodies used and reagents were: anti-SKP2 8D9 (Life Technologies); anti-MEF2C [35 (link)] anti-MEF2D α1/α2 [10 (link)]; anti-GFP, anti-HDAC4 [60 (link)] and anti-HDAC5 [38 (link)]; anti-HDAC7 (sc-74563), anti-MEF2A (C-21 sc-313) and anti-RACK1 (sc-17754, Santa Cruz Biotechnology); anti-MEF2D (BD Transduction Laboratories); anti-Actin, p21 CP74 and FLAG M2 (Sigma-Aldrich); anti-ubiquitin (Covance); anti-HDAC9 (ab109446), anti-H3K27ac (ab4729) and anti-H3K27me3 (ab6002) (Abcam); anti-H3K4me3 (GTX128954, GeneTex); anti-KI67 (556003, BD Pharmingen).
Anti rack1 sc 17754
Manufactured by Santa Cruz Biotechnology
Sourced in United States, United Kingdom
Anti-RACK1 (sc-17754) is a primary antibody that binds to the RACK1 protein. RACK1 is a scaffolding protein involved in various cellular processes. The antibody can be used to detect and analyze the RACK1 protein in samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti mef2d, by Santa Cruz Biotechnology (1 mentions)
Anti mef2d, by BD (1 mentions)
Anti ubiquitin, by Fortrea (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti erk, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
Cortisol, by Merck Group (1 mentions)
Mifepristone, by Merck Group (1 mentions)
Anti lamin a c 612162, by BD (1 mentions)
Host specific peroxidase conjugated igg secondary antibodies, by Thermo Fisher Scientific (1 mentions)
Electrophoresis reagents, by Bio-Rad (1 mentions)
Anti β tubulin t0198, by Merck Group (1 mentions)
Cell culture media, by Merck Group (1 mentions)
4 protocols using anti rack1 sc 17754
1
Antibodies for Molecular Biology Assays
2
Pharmacological Inhibition of Protein Kinase
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Calphostin C (Calbiochem, Merck KGaA, Darmstadt, Germany) was dissolved in ethanol to produce a 100 μM stock solution. Calphostin C was added to achieve a final concentration of 3 μM for 2 h before starting experiments. Hispidin from Calbiochem (Merck KGaA, Germany) was dissolved in DMSO to get a 10 mM stock solution. Hispidin was added to the culture cells at a concentration of 5 μM for 30 min. Anti-RACK1 (sc-17754, Santa Cruz Biotechnology, Dallas, TX, USA) was applied in the internal solution of the micropipette at a concentration of 0.4 µg/µL.
3
Western Blot Quantification of Signaling Proteins
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Western blot analysis was performed as previously described [26 (link)]. The primary antibodies used in this study were anti-CPNE1 (ab155675) (Abcam, UK), anti-RACK1 (sc-17754) (Santa Cruz, USA), anti-pEGFR (Tyr1068) (1H12) (#3777), anti-EGFR (#4267), anti-pAKT (Ser473) (D9E) (#4060), anti-AKT (#4685), anti-pERK (Thr202/Tyr202) (D13.14.4E) (#4370), anti-ERK (#4695), and anti-β-actin (#3700) (Cell Signaling Technology, USA). The bands were developed by an electrochemiluminescence reagent, imaged with a ChemiDoc XRS + (Bio-Rad, USA), and finally quantified with ImageJ software (National Institutes of Health, Bethesda, MD, USA).
4
Cortisol and Mifepristone Protocol
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Cortisol (PubChem CID:5754) and mifepristone (PubChem CID:55245) were obtained from Sigma Aldrich (St. Louis, MO, USA). They were dissolved in DMSO at concentration of 1 mM and 10 mM and frozen (−20 °C) in stock aliquots. Stock aliquots were diluted at a final concentration in culture media at the time of use (final concentration of DMSO in culture medium <0.1 %). Cell culture media and all supplements were obtained from Sigma Aldrich. Anti-RACK1 (sc17754) and anti-SRp30c (sc-134036) were obtained from Santa Cruz Biotechnology. Primers and probes of Grα, Grβ, Rack1, total Bdnf and 18S were purchased by Eurofins genomics whereas Bdnf long 3′UTR and Bdnf isoform IV were bought by Life technologies Italia. Anti-BDNF (SAB 1405514) and anti β-tubulin (T0198) were purchased from Sigma Aldrich. Anti lamin A/C (612162) was obtained from BD Biosciences (Franklin Lakes, NJ, USA). Host-specific peroxidase-conjugated IgG secondary antibodies were purchased from Thermo Scientific Inc. (Waltham,MA, USA). Electrophoresis reagents were purchased from Bio-Rad (Richmond, CA, USA).
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